Tumor hypoxia directed multimodal nanotherapy for overcoming drug resistance in renal cell carcinoma and reprogramming macrophages

Abstract

Drug resistance is one of the significant clinical burden in renal cell carcinoma (RCC). The development of drug resistance is attributed to many factors, including impairment of apoptosis, elevation of carbonic anhydrase IX (CA IX, a marker of tumor hypoxia), and infiltration of tumorigenic immune cells. To alleviate the drug resistance, we have used Sorafenib (Sor) in combination with tumor hypoxia directed nanoparticle (NP) loaded with a new class of apoptosis inducer, CFM 4.16 (C4.16), namely CA IX-C4.16. The NP is designed to selectively deliver the payload to the hypoxic tumor (core), provoke superior cell death in parental (WT) and Everolimus-resistant (Evr-res) RCC and selectively downmodulate tumorigenic M2-macrophage. Copper-free 'click' chemistry was utilized for conjugating SMA-TPGS with Acetazolamide (ATZ, a CA IX-specific targeting ligand). The NP was further tagged with a clinically approved NIR dye (S0456) for evaluating hypoxic tumor core penetration and organ distribution. Imaging of tumor spheroid treated with NIR dye-labeled CA IX-SMA-TPGS revealed remarkable tumor core penetration that was modulated by CA IX-mediated targeting in hypoxic-A498 RCC cells. The significant cell killing effect with synergistic combination index (CI) of CA IX-C4.16 and Sor treatment suggests efficient reversal of Evr-resistance in A498 cells. The CA IX directed nanoplatform in combination with Sor has shown multiple benefits in overcoming drug resistance through (i) inhibition of p-AKT, (ii) upregulation of tumoricidal M1 macrophages resulting in induction of caspase 3/7 mediated apoptosis of Evr-res A498 cells in macrophage-RCC co-culturing condition, (iii) significant in vitro and in vivo Evr-res A498 tumor growth inhibition as compared to individual therapy, and (iv) untraceable liver and kidney toxicity in mice. Near-infrared (NIR) imaging of CA IX-SMA-TPGS-S0456 in Evr-res A498 RCC model exhibited significant accumulation of CA IX-oligomer in tumor core with >3-fold higher tumor uptake as compared to control. In conclusion, this proof-of-concept study demonstrates versatile tumor hypoxia directed nanoplatform that can work in synergy with existing drugs for reversing drug-resistance in RCC accompanied with re-education of tumor-associated macrophages, that could be applied universally for several hypoxic tumors.

PDL-1 Antibody Drug Conjugate for Selective Chemo-Guided Immune Modulation of Cancer

“…From Figure 4F, it can be seen that the IFN-γ production in PDL1-Dox treatment is significantly higher compared to Dox in a co-cultured condition of MDA-MB-231 and activated RAW 264.7 cells. Literature reports indicate that activation of Raw 264.7 (macrophage) cells with lipopolysaccharide (LPS) can significantly upregulate the PD-1 expression [25,26]. Towards this end, we have utilized the LPS activated Raw 265.7 cells co-cultured with MDA-MB-231 and found the up-modulation of IFN-γ, suggesting the PDL1-Dox mediated inhibition of PD1 and PDL1 interaction.…”

Molecular Repolarisation of Tumour-Associated Macrophages

“…This nanoparticle delivery subsequently reprogrammed macrophages to an M1-like state in vitro, via inhibition of AKT1 phosphorylation preventing the activation of STAT6. Nanoparticle-treatment potently inhibited tumour growth in a mouse model of renal cell carcinoma [109]. Similarly, Zhang et al recently reported on the combination of sorafinib with HDAC inhibitor panobinostat and bromodomain protein inhibitor OTX015.…”

Progress in Clinical Trials of Photodynamic Therapy for Solid Tumors and the Role of Nanomedicine

“…In this context, nanocarriers need to provide sufficient therapeutic efficacy, and issues such as unfavorable biodistribution or rapid drug clearance from tumor areas must be resolved [ 81 ]. Furthermore, nanocarriers will experience a prolonged presence in the blood flow, aggregate in the tumor microenvironment, and facilitate efficient drug uptake by cancer cells [ 82 ].…”